WO2021244051A1 - 一种11-o-罗汉果醇肟醚衍生物及其制备方法 - Google Patents

一种11-o-罗汉果醇肟醚衍生物及其制备方法 Download PDF

Info

Publication number
WO2021244051A1
WO2021244051A1 PCT/CN2021/074164 CN2021074164W WO2021244051A1 WO 2021244051 A1 WO2021244051 A1 WO 2021244051A1 CN 2021074164 W CN2021074164 W CN 2021074164W WO 2021244051 A1 WO2021244051 A1 WO 2021244051A1
Authority
WO
WIPO (PCT)
Prior art keywords
oxime
group
ether derivative
mogrosteol
oxime ether
Prior art date
Application number
PCT/CN2021/074164
Other languages
English (en)
French (fr)
Inventor
李伟
黄华学
黄�俊
贺进军
宋谷良
Original Assignee
湖南华诚生物资源股份有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 湖南华诚生物资源股份有限公司 filed Critical 湖南华诚生物资源股份有限公司
Publication of WO2021244051A1 publication Critical patent/WO2021244051A1/zh

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J9/00—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/52—Adding ingredients
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/575—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of three or more carbon atoms, e.g. cholane, cholestane, ergosterol, sitosterol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Definitions

  • the present invention relates to 11-O-mogrosteol derivatives and preparation methods thereof, and in particular to 11-O-mogroscopy oxime ether derivatives and preparation methods thereof.
  • Mogroside is a unique medicinal and edible plant in my country, rich in various nutrient elements, flavonoids, polysaccharides, polyphenols and other biologically active substances.
  • Mogroside especially Mogroside V, is currently the most popular sweetener to replace sucrose. It is popular in European and American markets for its high sweetness and low calorie characteristics.
  • the carbon atoms on the saponin nucleus can form carbon radicals, which are further oxidized to form peroxy radicals.
  • the aglycone and hydroxyl group of tetracyclic triterpene saponins are the key groups with antioxidant activity. However, the antioxidant mechanism of mogroside saponins has not been studied in depth, and further research is needed.
  • Mogrol is the aglycon of mogroside IA1, IE1, IIE, III, IV, IVE, V and siamenoside I; 11-O-Mogrol (11-O-Mogrol), also known as 11- Oxime-mogroside is the aglycon of 11-O-mogroside V.
  • mogroside can significantly improve senile dementia (Alzheimer's disease, AD) and ischemic dementia-induced spatial cognitive impairment and decline in learning and memory.
  • Mogroside can increase the content of inhibitory amino acids, glycine and ⁇ -aminobutyric acid in the cerebral cortex of Alzheimer’s rats, reduce the toxicity of excitatory glutamate, and reduce the production of ⁇ -AP in the hippocampus of AD rats. It also has a significant improvement effect on the decline of learning and memory function caused by cerebral ischemia.
  • Mogroside can also significantly improve memory impairment in mice caused by NaNO 2. It can be seen that mogroside has obvious anti-senile dementia effects and can be used as a clinical drug for development and use.
  • the compounds with the oxime ether structure have aroused the research interest of scientists due to their unique structural characteristics and unique physiological activities in biochemistry.
  • Some of the compounds with oxime ether structure are related to the biological endocrine system, and some have the effects of antibacterial and anti-inflammatory, anti-oxidation, insecticide and weed control, and are also low-toxic and environmentally friendly. They can be widely used in biomedicine and biological pesticides. , Health food and medicine, so it has a good development prospect. Therefore, this patent discloses a 11-O-mogroscopy ether derivative (also called 11-oxime ether-mogroscopy derivative) and a preparation method thereof.
  • the present invention explores and determines the synthetic method of 11-O-mogrosteol oxime ether derivative and the biological activity of anti-oxidation.
  • the present invention discloses 11-O-mogrosteol oxime ether derivative (also can be called 11-oxime ether-mogrosteol derivative) and a preparation method thereof.
  • the specific technical solutions are as follows:
  • R is H, alkyl, alkoxy, alkenyl, alkynyl, alkylene-aryl, alkylene-heteroaryl.
  • the alkyl group, alkoxy group, alkenyl group, and alkynyl group have 1-20 carbon atoms, preferably 1-6 carbon atoms; the alkylene group has 1-3 carbon atoms; the aryl group, hetero The aryl group has 4-20 carbon atoms, preferably 5-12 carbon atoms, the heteroaryl group also has 1-3 heteroatoms, and the heteroatoms are selected from O, S, N, and P.
  • the alkyl group is selected from C1-6 alkyl groups, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl;
  • the alkenyl group is selected from vinyl, Propenyl, butenyl;
  • the alkynyl group is selected from ethynyl, propynyl, butynyl;
  • the aryl group is selected from phenyl, naphthyl;
  • the heteroaryl group is selected from furyl, piperidinyl furanyl , Thienyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, oxazolyl, 1,3,4-oxadiazolyl, thiazolyl, benzimidazole, benzothiophene, benzofuran.
  • the present invention provides the following specific compounds:
  • 11-O-mogroscopy oxime ether derivatives have stronger antioxidant activity and better water solubility, which is more conducive to human body absorption and utilization, especially when taken orally.
  • the utilization rate is significantly improved.
  • the present invention also provides a preparation method of the 11-O-mogrosteol oxime ether derivative, which comprises the following steps:
  • Oximation Dissolve 11-O-Mogrostil in alcohol, add hydroxylamine salt and acid binding agent, reflux reaction, after the reaction, concentrate under reduced pressure, cool the concentrated solution, stir to crystallize, filter, and dry to obtain 11-oxime -Momordinol;
  • the synthetic route of the 11-O-Mogrosteol oxime ether derivative is as follows:
  • the 11-O-mogroside is derived from the aglycon obtained after the hydrolysis of the natural ingredient 11-O-mogroside V, or is semi-synthesized or fully synthesized from mogroside.
  • the alcohol is anhydrous methanol or anhydrous ethanol, and the amount of alcohol is 4-20 times (ml/g) of the weight of 11-O-mogrosteol.
  • the hydroxylamine salt is hydroxylamine hydrochloride, hydroxylamine sulfate or hydroxylamine phosphate, wherein the molar ratio of the hydroxylamine group to the 11-O-mogroscopy is (1.0-1.5):1.
  • the acid binding agent is an organic base, specifically pyridine, piperidine, imidazole, methylimidazole, dimethylamine, diethylamine, trimethylamine, triethylamine, ethylenediamine
  • the molar ratio of the acid binding agent to 11-O-mogroscopy is (0.5-5):1.
  • the reaction temperature is 65-85°C, and the reaction time is 2-10 hours.
  • the solid content in the concentrated solution is 20%-30%
  • the cooling temperature is 5-10°C
  • the stirring speed is 10-20r/min
  • the crystallization time is 12-24 hours.
  • the dosage of the DMF is 5-10 times (ml/g) of the weight of 11-oxime-mogroscopy.
  • the halogenating reagent is a saturated halogenated alkane, an unsaturated halogenated alkane or a halogenated aromatic hydrocarbon, and the molar ratio of the halogenating reagent to 11-oxime-mohangol is (1.0-1.05):1.
  • the catalyst is sodium hydride, potassium methoxide, sodium methoxide, potassium ethoxide, sodium ethoxide, potassium tert-butoxide, sodium tert-butoxide, sodium hydroxide, sodium carbonate, potassium carbonate, carbonic acid Ammonium, benzyl tetradecyl dimethyl ammonium chloride or tetrabutyl ammonium iodide, the amount of the catalyst used is 0.1% to 2% of the amount (mol) of the 11-oxime-mogroscopy substance.
  • the reaction temperature is -10 to 10°C, and the reaction time is 2 to 10 hours.
  • the amount of petroleum ether is 20-80 times (ml/g) of the weight of 11-oxime-mogroscopy.
  • the purpose of petroleum ether extraction is to transfer the product of the etherification reaction from the water phase to the petroleum ether phase to facilitate subsequent silica gel column chromatography separation.
  • the silica gel is column chromatography silica gel
  • the specification of the silica gel is 60-300 mesh
  • the amount of silica gel is 10-40 times the weight of 11-oxime-mogrostil
  • the silica gel column The ratio of height to diameter is (2 ⁇ 8):1.
  • the purpose of using silica gel chromatography column is to adsorb and separate high content of 11-oxime ether-mogrosultol.
  • the flow rate of the petroleum ether layer through the silica gel chromatography column is 0.5-1 BV/hour.
  • the eluent is a mixed solution of petroleum ether and ethyl acetate, dichloromethane or chloroform (any one of the three solutions), petroleum ether and ethyl acetate,
  • the volume ratio of methylene chloride or chloroform is (7-10):1, and the amount of the eluent is 2-5 BV.
  • the flow rate of the elution is 0.5-2 BV/hour.
  • 1BV 1 volume of silica gel chromatography column.
  • the present invention also provides the use of the 11-O-mogrosteol oxime ether derivative as an antioxidant, which is used as an additive in food, health care products, beverages, and medicines.
  • the 11-O-mogroside used in the examples of the present invention was purchased from Hunan Huacheng Biological Resources Co., Ltd. It is an aglycon obtained by hydrochloric acid hydrolysis from the natural ingredient of Luo Han Guo—11-O-mogroside V.
  • the purity measured by high performance liquid chromatography (HPLC) area normalization method was 97.5%, and the weight loss on drying and ash content of the 11-O-mogrosultol sample were measured to be less than 0.5%; the auxiliary materials or chemical reagents used in the examples of the present invention , Unless otherwise specified, all are obtained through conventional commercial channels.
  • Oximation and post-treatment take 23.8g of 11-O-mogroscinol and dissolve it in 120ml of absolute ethanol, add 4.17g of hydroxylamine hydrochloride (containing 0.06mol of hydroxylamine) and 7.9g of pyridine, reflux at 82°C for 4 hours, silica gel After the completion of the TLC inspection reaction, the concentrated solution was concentrated under reduced pressure to 20% of the solid content of the concentrated solution. The concentrated solution was cooled to a temperature of 5°C, stirred at a speed of 10r/min, crystallized for 20 hours, filtered with suction, and dried to obtain 11-oxime- Mogroside is 22.1g.
  • Oximation and post-treatment Take 23.8g of 11-O-Mogroscarol and dissolve it in 150ml of anhydrous methanol, add 4.5g of hydroxylamine sulfate (containing 0.055mol of hydroxylamine) and 20.2g of triethylamine, and react at 70°C for 5 hours. After the silica gel TLC inspection reaction, it was concentrated under reduced pressure to 25% of the solid content of the concentrated liquid. The concentrated liquid was cooled to a temperature of 10°C, stirred at a speed of 10r/min, crystallized for 18 hours, filtered with suction, and dried to obtain 11- The oxime-mogroscinol 21.6g.
  • Oximation and post-treatment take 23.8g of 11-O-mogrosultol, dissolve it in 150ml of absolute ethanol, add 3.94g of hydroxylamine phosphate (containing 0.06mol of hydroxylamine) and 11g of diethylamine, reflux at 85°C for 6 hours, After the silica gel TLC inspection reaction, it was concentrated under reduced pressure until the solid content of the concentrated liquid was 22%. The concentrated liquid was cooled to a temperature of 7°C, stirred at a speed of 15r/min, crystallized for 16 hours, filtered with suction, and dried to obtain 11-oxime -Momordinol 20.6g.
  • mice were used, weighing 20g ⁇ 2g, half male and half male.
  • the 11-O-mogrosteol oxime ether derivative prepared in Example 1-3 was made into a 5% water suspension, and fasted mice were given intragastrically with 0.5 mL each time 3 times a day, and observation was continued for 7 days , No deaths, no toxic side effects.
  • Kunming mice were used, weighing 20g ⁇ 2g, half male and half male.
  • the 11-O-mogrosteol oxime ether derivative prepared in Examples 1-3 was formulated into a 3% injection. Administration by injection, 0.4 mL each time. Observed for 7 consecutive days, there was no death or toxicity.
  • the 11-O-Mogrosteol oxime ether derivative prepared in Example 1-3 was formulated into a 10% aqueous suspension, and the dosage was 5 mg/kg/d, 10 mg/kg/d, and 20 mg/kg every two weeks. d.
  • the six concentrations of 40mg/kg/d, 60mg/kg/d, and 80mg/kg/d are gradually increased, and the drug is continuously administered every day for 12 weeks. There was no significant difference in peripheral blood, liver, kidney function, blood sugar and other biochemical indicators; no significant pathological improvement was caused to important organs, indicating that there was no obvious toxic and side effects at 80 mg/kg/d.
  • Free radical DPPH scavenging test the DPPH method was used to quantitatively determine the antioxidant capacity of 11-O-mogroside oxime ether derivatives. The principle is that DPPH free radicals have a single electron, which has a strong absorption at 520nm, and the alcohol solution is purple. Antioxidants can make one electron pair, so that the A520nm value is reduced and the solution fades. Since this change is a quantitative relationship with the number of electrons accepted, it can be measured with a colorimetric method (such as a spectrophotometer). Proceed as follows:
  • test product Take the 11-O-mogrosteol oxime ether derivative sample obtained in the example, add 95% ethanol solution to constant volume, and prepare 100 ⁇ g/mL test solution respectively.
  • the 11-O-mogrosultol control group and vitamin C group were also distributed and prepared at the concentration of 100 ⁇ g/mL.
  • Inhibition rate (%) ( ⁇ A0- ⁇ A)/ ⁇ A0 ⁇ 100 where: ⁇ A0 is the auto-oxidation rate of pyrogallol; ⁇ A is the auto-oxidation rate of pyrogallol after adding the sample solution.
  • Test dogs 40 dogs, ten in each group, half male and half male, 10 ⁇ 1kg, purchased from Hunan Provincial Animal Experimental Center.
  • the sample was prepared as a 50 mg/mL sterile aqueous solution and administered to dogs in a fasted state through an oral feeding tube method. Each group of ten dogs, half of the male and the female, was administered at a dose of 10 mg/kg. After the administration, blood was collected at the same site at each time period of 0min, 15min, 30min, 45min, 1h, 1.5h, 2h, 4h, 6h, 8h, and 10h, and the plasma was separated for testing. The concentration of the 11-O-mogrosteol oxime ether derivative of the present invention in dog plasma is tested by HPLC/MS method. T1/2 represents the half-life of the drug.
  • the data in Table 1 show that the 11-O-Mogrostilbene oxime ether derivative provided by the present invention has significantly enhanced antioxidant activity and solubility compared to the parent compound 11-O-Mogrostilbene, and is a new type to be developed and studied.
  • Antioxidants for natural products have an activity close to that of commonly used antioxidants Vc, and their half-life in organisms is significantly prolonged. They are suitable for use as antioxidant additives in oral products, such as foods, beverages, health products, and dairy products.

Abstract

本发明提供了一种11-O-罗汉果醇肟醚衍生物及其制备方法，结构式如式(I)所示。所述罗汉果醇肟醚衍生物是在11-O-罗汉果醇的酮基肟化后再进行醚化，得到产物11-O-罗汉果醇肟醚衍生物。本发明提供的11-O-罗汉果醇肟醚衍生物制备方法简单，纯度和产率高。所得11-O-罗汉果醇肟醚衍生物相比于11-O-罗汉果醇，具有明显增强的抗氧化活性、水溶性和体内半衰期，而且没有毒性，可以作为食品，饮品，保健品，药品等口服产品中的抗氧化功能添加剂。

Description

一种11-O-罗汉果醇肟醚衍生物及其制备方法 技术领域
本发明涉及11-O-罗汉果醇衍生物及其制备方法，具体涉及11-O-罗汉果醇肟醚衍生物及其制备方法。
背景技术
罗汉果是我国独有的药食同源植物，富含各种营养元素和黄酮，多糖，多酚等生物活性物质。罗汉果苷，特别是罗汉果甜苷V是目前最受欢迎的代替蔗糖的甜味剂，以高甜度，低热量的特点在欧美市场备受欢迎。有文献对罗汉果苷的生物活性特别是抗氧化作用进行了研究，发现罗汉果苷V具有一定的抗氧化能力，对羟基和超氧阴离子自由基均有一定的清除作用。一般认为罗汉果苷具有抗氧化活性是因为苷元上氢原子有一定活性，可以和羟基自由基结合，皂苷母核上的碳原子可以形成碳自由基，进一步氧化形成过氧自由基。四环三萜类皂苷的苷元以及羟基是具有抗氧化活性的关键集团。但罗汉果皂苷类化合物的抗氧化机制还没有深入研究，有待进一步探究。
罗汉果醇(Mogrol)是罗汉果甜苷ⅠA1、ⅠE1、ⅡE、Ⅲ、Ⅳ、ⅣE、Ⅴ和赛门苷I等的苷元；11-O-罗汉果醇(11-O-Mogrol)，也即11-肟-罗汉果醇，则是11-O-罗汉果苷V的苷元。
现代科学研究发现，罗汉果醇对于老年性痴呆(阿尔茨海默症，AD)以及缺血性脑痴呆引起的空间认知障碍和学习记忆功能下降具有明显的改善作用。罗汉果醇能够增加老年性痴呆大鼠大脑皮层抑制性氨基酸、甘氨酸、γ-氨基丁酸的含量，降低兴奋性谷氨酸的毒性，并能减少AD大鼠大脑海马组织中β-AP的产生，对于脑缺血引起的学习记忆功能下降也具有明显改善作用。罗汉果醇也能明显改善NaNO 2所致的小鼠记忆障碍。由此可见，罗汉果醇具有明显的抗老年痴呆的作用，可以作为临床药物开发使用。
近年来，具有肟醚结构的化合物因其独特的结构特征，以及在生物化学上特有的生理活性，已经引起了科学工作者的研究兴趣。具有肟醚结构的化合物有的与生物内分泌系统相关，有的具有抗菌消炎、抗氧化、杀虫除草等作用，而且还有低毒和环境友好的特点，可以广泛的应用到生物医药、生物农药、保健食品和药品中，因此具有很好的发展前景。因此，本专利公开一种11-O-罗汉果醇肟醚衍生物(也可以叫做11-肟醚-罗汉果 醇衍生物)及其制备方法。
目前，尚没有11-O-罗汉果醇肟醚衍生物的相关报道。本发明探究并确定了11-O-罗汉果醇肟醚衍生物的合成方法以及抗氧化的生物活性。
发明内容
本发明解决其技术问题所采用的技术方案如下：
本发明公开了11-O-罗汉果醇肟醚衍生物(也可以叫做11-肟醚-罗汉果醇衍生物)及其制备方法。具体技术方案如下：
一种11-O-罗汉果醇肟醚衍生物，结构式如下式(I)所示：
Figure PCTCN2021074164-appb-000001
其中，R为H、烷基、烷氧基、烯基、炔基、亚烷基-芳基、亚烷基-杂芳基。
所述烷基、烷氧基、烯基、炔基具有1-20个碳原子，优选具有1-6个碳原子；所述亚烷基具有1-3个碳原子；所述芳基，杂芳基具有4-20个碳原子，优选具有5-12个碳原子，所述杂芳基还有1-3个杂原子，所述杂原子选自O，S，N，P。
优选地，所述烷基选自C1-6的烷基，比如甲基、乙基、丙基、异丙基、丁基、异丁基、叔丁基；所述烯基选自乙烯基、丙烯基、丁烯基；所述炔基选自乙炔基，丙炔基，丁炔基；所述芳基选自苯基，萘基；所述杂芳基选自呋喃基，哌啶呋喃基、噻吩基、吡咯基、吡唑基、咪唑基、三唑基、噁唑基、1，3，4-噁二唑基、噻唑基、苯并咪唑，苯并噻吩，苯并呋喃。
更为优选地，本发明提供以下具体化合物：
Figure PCTCN2021074164-appb-000002
发明人发现，相比于11-O-罗汉果醇，11-O-罗汉果醇肟醚衍生物有着更强的抗氧化活性和更好的水溶性，更有利于人体吸收利用，特别是口服，生物利用率显著提高。
本发明还提供了所述11-O-罗汉果醇肟醚衍生物的制备方法，包括以下步骤：
(1)肟化：将11-O-罗汉果醇溶解于醇，加入羟胺盐以及缚酸剂，回流反应，反应结束后减压浓缩，浓缩液冷却，搅拌结晶，过滤，干燥，得11-肟-罗汉果醇；
(2)醚化：将11-肟-罗汉果醇溶解于DMF，加入卤化试剂以及催化剂，低温反应，反应结束后将反应液倒入冰水中，石油醚萃取，收集石油醚层，用水将石油醚层洗至中性，将石油醚层通过硅胶层析柱，用洗脱剂洗脱，收集洗脱液，浓缩，挥干溶剂，得11-肟醚-罗汉果醇。
所述11-O-罗汉果醇肟醚衍生物的合成线路如下：
Figure PCTCN2021074164-appb-000003
优选地，步骤(1)中，所述的11-O-罗汉果醇来源于天然成分11-O-罗汉果苷V水解后得到的苷元，或者由罗汉果醇半合成，或者全合成。
优选地，步骤(1)中，所述的醇为无水甲醇或无水乙醇，醇的用量为11-O-罗汉 果醇重量的4～20倍(ml/g)。
优选地，步骤(1)中，所述的羟胺盐为盐酸羟胺、硫酸羟胺或磷酸羟胺，其中羟胺基团与11-O-罗汉果醇的摩尔比为(1.0～1.5)：1。
优选地，步骤(1)中，所述的缚酸剂为有机碱，具体是指吡啶、哌啶、咪唑、甲基咪唑、二甲胺、二乙胺、三甲胺、三乙胺、乙二胺、丙二胺、环己胺、二环己胺中的一种或几种，所述缚酸剂与11-O-罗汉果醇的摩尔比为(0.5～5)：1。优选地，步骤(1)中，所述的反应温度为65～85℃，反应的时间为2～10小时。
优选地，步骤(1)中，所述的浓缩液中固形物含量为20％～30％，所述冷却的温度为5～10℃，所述搅拌的速度为10～20r/min，所述析晶的时间为12～24小时。
优选地，步骤(2)中，所述DMF的用量为11-肟-罗汉果醇重量的5～10倍(ml/g)。
优选地，步骤(2)中，所述的卤化试剂为饱和卤代烷、不饱和卤代烷或卤代芳香烃，所述卤化试剂与11-肟-罗汉果醇的摩尔比为(1.0～1.05)：1。
优选地，步骤(2)中，所述的催化剂为氢化钠、甲醇钾、甲醇钠、乙醇钾、乙醇钠、叔丁醇钾、叔丁醇钠、氢氧化钠、碳酸钠、碳酸钾、碳酸铵、苄基十四烷二甲基氯化铵或四丁基碘化铵，所述催化剂的用量为11-肟-罗汉果醇物质的量(mol)的0.1％～2％。
优选地，步骤(2)中，所述的反应温度为-10～10℃，反应的时间为2～10小时。
优选地，步骤(2)中，所述石油醚的的用量为11-肟-罗汉果醇重量的20～80倍(ml/g)。石油醚萃取的目的是将醚化反应的产物从水相转移至石油醚相，便于后续硅胶柱层析分离。
优选地，步骤(2)中，所述的硅胶为柱层析硅胶，硅胶的规格为60～300目，硅胶的用量为11-肟-罗汉果醇重量的10～40倍，所述硅胶柱的高径比为(2～8)：1。使用硅胶层析柱的目的吸附并分离高含量的11-肟醚-罗汉果醇。
优选地，步骤(2)中，所述石油醚层通过硅胶层析柱的流速为0.5～1BV/小时。
优选地，步骤(2)中，所述的洗脱剂为石油醚与乙酸乙酯、二氯甲烷或三氯甲烷(三者中任意一种溶液)的混合溶液，石油醚与乙酸乙酯、二氯甲烷或三氯甲烷的体积比为(7～10)：1，所述洗脱剂的用量为2～5BV。
优选地，步骤(2)中，所述洗脱的流速为0.5～2BV/小时。
本发明方法中，1BV＝1个硅胶层析柱体积。
本发明还提供了所述11-O-罗汉果醇肟醚衍生物作为抗氧化剂的用途，用于食品， 保健品，饮品，药品中的添加剂。
具体实施方式
下面结合实施例对本发明作进一步说明。
本发明实施例所使用的11-O-罗汉果醇购于湖南华诚生物资源股份有限公司，是用罗汉果天然成分——11-O-罗汉果甜苷V——经盐酸水解得到的苷元，经高效液相色谱(HPLC)面积归一法测得纯度为97.5％，另测得该11-O-罗汉果醇样品的干燥失重、灰分均小于0.5％；本发明实施例所使用的辅料或化学试剂，如无特殊说明，均通过常规商业途径获得。
实施例1
(1)肟化、后处理：取11-O-罗汉果醇23.8g,溶解于120ml无水乙醇，加入盐酸羟胺4.17g(含羟胺0.06mol)以及吡啶7.9g，82℃回流反应4小时，硅胶TLC检验反应结束后，减压浓缩至浓缩液固形物含量20％，将浓缩液冷却至温度5℃，以10r/min的速度搅拌，析晶20小时，抽滤，干燥，得11-肟-罗汉果醇22.1g。
(2)醚化、后处理：取步骤(1)所得11-肟-罗汉果醇20.2g溶解于110ml DMF，加入溴乙烷4.58g以及甲醇钠0.022g，0℃反应6小时，硅胶TLC检验反应结束后，将反应液倒入1000ml冰水中，用500ml石油醚萃取，收集石油醚层，用水将石油醚层洗至中性，将石油醚层以0.7BV/小时的流速通过硅胶层析柱(柱层析硅胶的规格为100～200目，硅胶的用量为300g，硅胶层析柱的高径比为6：1)，用5BV洗脱剂(石油醚：乙酸乙酯＝9：1，V/V)洗脱，洗脱的流速为1BV/小时，收集洗脱液，浓缩，挥干溶剂，得11-肟乙醚-罗汉果醇18.7g，纯度99.4％，产率72％，其结构如式(I-1)所示。
1H NMR(CDCl 3)：1.05(d,3H)，1.13-1.21(m,15H),1.24-1.26(m,11H),1.29(q,2H)，1,43-1.47(m,5H),1.65(t，1H)，1.92(d,2H)1.97(t,1H),3.23(t，1H)，3.28(t,1H),3.58(q,2H),5.37(s,1H)。Anal.Calcd.for C 32H 55NO 4:C.74.22,H.10.72,N.2.71,O.12.35。m/z:517.41,Found:517.73。
实施例2
(1)肟化、后处理：取11-O-罗汉果醇23.8g,溶解于150ml无水甲醇，加入硫酸羟胺4.5g(含羟胺0.055mol)以及三乙胺20.2g，70℃回流反应5小时，硅胶TLC检验反应结束后，减压浓缩至浓缩液固形物含量25％，将浓缩液冷却至温度10℃，以10r/min的速度搅拌，析晶18小时，抽滤，干燥，得11-肟-罗汉果醇21.6g。
(2)醚化、后处理：取步骤(1)所得11-肟-罗汉果醇20.2g(0.04mol)溶解于110ml DMF，加入氯丙烯3.14g以及叔丁醇钠0.08g，5℃反应5小时，硅胶TLC检验反应结束后，将反应液倒入1000ml冰水中，用800ml石油醚萃取，收集石油醚层，用水将石油醚层洗至中性，将石油醚层以1BV/小时的流速通过硅胶层析柱(柱层析硅胶的规格为80～100目，硅胶的用量为400g，硅胶层析柱的高径比为5：1)，用3BV洗脱剂(石油醚：二氯甲烷＝7：1，V/V)洗脱，洗脱的流速为1.5BV/小时，收集洗脱液，浓缩，挥干溶剂，得11-肟丙烯醚-罗汉果醇17.4g，纯度99.1％，产率65.7％，其结构如式(I-2)所示。
1H NMR(CDCl 3)：1.05(d,3H)，1.13-1.21(m,12H),1.24-1.26(m,11H),1.29(q,2H)，1,43-1.47(m,5H),1.65(t，1H)，1.92(d,2H)1.97(t,1H),3.23(t，1H)，3.28(t,1H),4.22(d,2H),5.23(d,2H),，5,37(s,1H)，5.92(s,1H)。Anal.Calcd.for C 33H 55NO 4:C.74.81,H.10.45,N.2.64,O.12.09。m/z:529.38,Found:529.56。
实施例3
(1)肟化、后处理：取11-O-罗汉果醇23.8g,溶解于150ml无水乙醇，加入磷酸羟胺3.94g(含羟胺0.06mol)以及二乙胺11g，85℃回流反应6小时，硅胶TLC检验反应结束后，减压浓缩至浓缩液固形物含量22％，将浓缩液冷却至温度7℃，以15r/min的速度搅拌，析晶16小时，抽滤，干燥，得11-肟-罗汉果醇20.6g。
(2)醚化、后处理：取步骤(1)所得11-肟-罗汉果醇20.2g溶解于120ml DMF，加入溴化苄6.84g以及四丁基碘化铵0.3g，-3℃反应8小时，硅胶TLC检验反应结束后，将反应液倒入1000ml冰水中，用1000ml石油醚萃取，收集石油醚层，用水将石油醚层洗至中性，将石油醚层以0.5BV/小时的流速通过硅胶层析柱(柱层析硅胶的规格为200～300目，硅胶的用量为230g，硅胶层析柱的高径比为8：1)，用4BV洗脱剂(石油醚：三氯甲烷＝9.5：1，V/V)洗脱，洗脱的流速为0.7BV/小时，收集洗脱液，浓缩，挥干溶剂，得11-肟苄基醚-罗汉果醇15.7g，纯度98.7％，产率54.2％，其结构如式(I-3)所示。
1H NMR(CDCl 3)：1.05(d,3H)，1.12-1.19(m,12H),1.24-1.26(m,11H),1.29(q,2H)，1,43-1.47(m,5H),1.65(t，1H)，1.92(d,2H)1.97(t,1H),3.23(t，1H)，3.28(t,1H),4.82(s,2H),5.38(s,1H)，7.22-7.24(m，5H)。Anal.Calcd.for C 37H 57NO 4:C.76.62,H.9.92,N.2.42,O.11.04。m/z:579.64,Found:579.82。
效果例1  毒性实验
(1)口服给药
用昆明种小鼠，体重20g±2g，雌雄各半。将实施例1-3制备得到的11-O-罗汉果醇肟醚衍生物制成5％的水混悬液，一日3次对禁食小鼠灌胃、每次0.5mL，连续观察7天，未见死亡，未见毒副反应。
(2)注射给药
用昆明种小鼠，体重20g±2g，雌雄各半。将实施例1-3制备得到的11-O-罗汉果醇肟醚衍生物配制成3％的注射液。注射给药，每次0.4mL。连续观察7天，未见死亡，未见毒副反应。
(3)长期毒性试验
用SD大鼠，体重200±20g，雌雄各半，共180只。分别灌胃给药。将实施例1-3制备得到的11-O-罗汉果醇肟醚衍生物配制成10％的水混悬液，剂量按照每两周5mg/kg/d、10mg/kg/d、20mg/kg/d、40mg/kg/d，60mg/kg/d，80mg/kg/d六个浓度递增，每天连续给药，持续12周。对外周血象和肝，肾功能及血糖等生化指标均无明显差异；对重要脏器未引起明显的病理改善，表明80mg/kg/d时没有明显的毒副反应。
效果例2  抗氧化性的测试
1. 自由基DPPH的清除试验：采用DPPH法定量测定11-O-罗汉果醇肟醚衍生物抗氧化能力。原理是根据DPPH自由基有单电子，在520nm处有一强吸收，其醇溶液呈紫色。抗氧化剂能使单电子配对，从而使A520nm值降低，溶液褪色。由于这种变化其接受电子数量成定量关系，因而可用比色法(如分光光度计)进行测量。步骤如下：
(1)DPPH试剂的配制：精密称取DPPH(1,1-diphenyl-2-picrylhydrazyl)粉末(分子量＝394)0.00990g，置于250mL容量瓶，用适量95％乙醇溶解定容至250mL，得浓度为0.10mmol/L的DPPH试剂。
(2)供试品的制备：取实施例所得11-O-罗汉果醇肟醚衍生物样品，加入95％乙醇溶液定容，分别配制成100μg/mL试液。同样发放配制11-O-罗汉果醇对照组和维生素C组，浓度均为100μg/mL。
(3)测定方法：
a.在10ml试管中依次加入4.0ml DPPH溶液和1.0ml95％乙醇，混合摇匀，避光反应30min，稳定后，以95％乙醇为参比，在520nm处测吸光值，得空白管吸光值，记为A 0。
b.在10ml试管中依次加入4.0ml DPPH溶液和1.0ml待测试样溶液，混合摇匀，避 光反应30min，稳定后，以95％乙醇为参比，在520nm处测吸光值，得样品管吸光值，记为A 1。
c.在10ml试管中依次加入4.0ml95％乙醇溶液和1.0ml待测试样溶液，混合摇匀，避光反应30min，稳定后，以95％乙醇为参比，在520nm处测吸光值，得本底管吸光值，记为A 2。
d.计算公式：样品对DPPH自由基的清除率SA(％)：
Figure PCTCN2021074164-appb-000004
2. 超氧阴离子自由基(O 2-)的清除试验 ：邻苯三酚自氧化法。
采用邻苯三酚自氧化法，取4mL 0.1mol/L pH8.2Tris-HCl缓冲溶液和蒸馏水2mL，混匀后在25℃水浴中保温20min，然后加入样品溶液(100μg/mL)2mL，取出后立即加入在25℃预热过的5mmol/L邻苯三酚0.5mL(以10mmol/L HCL配制，空白管用10mmol/L HCL代替邻苯三酚的HCL溶液)，摇匀后倒入比色皿，325nm下每隔30s测定吸光度，连续测定4min，计算线性范围内每分钟吸光度的增加。在加入一定体积样品溶液时，减少蒸馏水的体积。
抑制率(％)＝(△A0-△A)/△A0×100式中：△A0为邻苯三酚的自氧化速率；△A为加入样品溶液后邻苯三酚的自氧化速率。
3. 水中溶解度采用HPLC外标法测试，所用水为去离子水，测试温度20℃。
4. 药代动力学。
试验用犬：40只，每组十只，雌雄各半，10±1kg，购自湖南省动物实验中心。
将样品配制为50mg/mL无菌水溶液，通过口饲管法给予禁食状态的狗，每组十只狗，雌雄各半，按照10mg/kg剂量给药。给药后，按照在0min，15min,30min，45min,1h，1.5h,2h，4h，6h，8h,10h每个时间段在相同部位采血，分离出血浆供测试用。本发明11-O-罗汉果醇肟醚衍生物在狗血浆中的浓度用HPLC/MS方法测试。T1/2表示药物半衰期。
所得数据结果如下表1所示：
表1
Figure PCTCN2021074164-appb-000005
表1数据表明，本发明提供的11-O-罗汉果醇肟醚衍生物相比于母体化合物11-O-罗汉果醇具有明显增强的抗氧化活性和溶解度，是一种有待开发和研究的新型来自于天然产物的抗氧化剂，其具有接近于常用抗氧化剂Vc的活性，而且在生物体内半衰期明显延长，适合作为口服产品，比如食品，饮品，保健品，乳制品等中的抗氧化添加剂。
最后应说明的是：以上各实施例仅用以说明本发明的技术方案，而非对其限制；尽管参照前述各实施例对本发明进行了详细的说明，本领域的普通技术人员应当理解：其依然可以对前述各实施例所记载的技术方案进行修改，或者对其中部分或者全部技术特征进行等同替换；而这些修改或者替换，并不使相应技术方案的本质脱离本发明各实施例技术方案的范围。

Claims (10)

  1. 一种11-O-罗汉果醇肟醚衍生物，结构式如下式(I)所示：
    Figure PCTCN2021074164-appb-100001
    其中，R为H、烷基、烷氧基、烯基、炔基、亚烷基-芳基、亚烷基-杂芳基。
  2. 如权利要求1所述的11-O-罗汉果醇肟醚衍生物，其特征在于，所述烷基、烷氧基、烯基、炔基具有1-20个碳原子，优选具有1-6个碳原子；所述亚烷基具有1-3个碳原子；所述芳基，杂芳基具有4-20个碳原子，优选具有5-12个碳原子，所述杂芳基还有1-3个杂原子，所述杂原子选自O，S，N，P。
  3. 如权利要求1所述的11-O-罗汉果醇肟醚衍生物，其特征在于，所述烷基选自C1-6的烷基，比如甲基、乙基、丙基、异丙基、丁基、异丁基、叔丁基；所述烯基选自乙烯基、丙烯基、丁烯基；所述炔基选自乙炔基，丙炔基，丁炔基；所述芳基选自苯基，萘基；所述杂芳基选自呋喃基，哌啶呋喃基、噻吩基、吡咯基、吡唑基、咪唑基、三唑基、噁唑基、1，3，4-噁二唑基、噻唑基、苯并咪唑，苯并噻吩，苯并呋喃。
  4. 如权利要求1所述的11-O-罗汉果醇肟醚衍生物，其特征在于，具有如下化学结构：
    Figure PCTCN2021074164-appb-100002
    Figure PCTCN2021074164-appb-100003
  5. 权利要求1-4任一项所述11-O-罗汉果醇肟醚衍生物的制备方法，其特征在于，合成路线如下：
    Figure PCTCN2021074164-appb-100004
  6. 权利要求1-4任一项所述11-O-罗汉果醇肟醚衍生物的制备方法，其特征在于，包括以下步骤：
    (1)肟化：将11-O-罗汉果醇溶解于醇，加入羟胺盐以及缚酸剂，回流反应，反应结束后减压浓缩，浓缩液冷却，搅拌结晶，过滤，干燥，得11-肟-罗汉果醇；
    (2)醚化：将11-肟-罗汉果醇溶解于DMF，加入卤化试剂以及催化剂，低温反应，反应结束后将反应液倒入冰水中，石油醚萃取，收集石油醚层，用水将石油醚层洗至中性，将石油醚层通过硅胶层析柱，用洗脱剂洗脱，收集洗脱液，浓缩，挥干溶剂，得11-肟醚-罗汉果醇。
  7. 如权利要求6所述的制备方法，其特征在于，步骤(1)中，所述的醇为无水甲醇或无水乙醇，醇的用量为11-O-罗汉果醇重量的4～20倍(ml/g)；和/或所述的羟胺盐为盐酸羟胺、硫酸羟胺、磷酸羟胺，其中羟胺基团与11-O-罗汉果醇的摩尔比为(1.0～1.5)：1；和/或所述缚酸剂与11-O-罗汉果醇的摩尔比为(0.5～5)：1。
  8. 如权利要求6所述的制备方法，其特征在于，步骤(1)中，所述的浓缩液中固形物含量为20％～30％，所述冷却的温度为5～10℃，所述搅拌的速度为10～20r/min，所述析晶的时间为12～24小时。
  9. 如权利要求6所述的制备方法，其特征在于，步骤(2)中，所述DMF的用量为11-肟-罗汉果醇重量的5～10倍(ml/g)；和/或所述的卤化试剂为饱和卤代烷、不饱 和卤代烷或卤代芳香烃，所述卤化试剂与11-肟-罗汉果醇的摩尔比为(1.0～1.05)：1；和/或所述的催化剂为氢化钠、甲醇钾、甲醇钠、乙醇钾、乙醇钠、叔丁醇钾、叔丁醇钠、氢氧化钠、碳酸钠、碳酸钾、碳酸铵、苄基十四烷二甲基氯化铵或四丁基碘化铵，所述催化剂的用量为11-肟-罗汉果醇物质的量(mol)的0.1％～2％。
  10. 权利要求1-4任一项所述11-O-罗汉果醇肟醚衍生物作为抗氧化剂的用途，其特征在于，用作食品、保健品、饮品、乳制品或药品中的抗氧化添加剂。
PCT/CN2021/074164 2020-06-01 2021-01-28 一种11-o-罗汉果醇肟醚衍生物及其制备方法 WO2021244051A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202010482693.8A CN111560044B (zh) 2020-06-01 2020-06-01 一种11-o-罗汉果醇肟醚衍生物及其制备方法
CN202010482693.8 2020-06-01

Publications (1)

Publication Number Publication Date
WO2021244051A1 true WO2021244051A1 (zh) 2021-12-09

Family

ID=72075138

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2021/074164 WO2021244051A1 (zh) 2020-06-01 2021-01-28 一种11-o-罗汉果醇肟醚衍生物及其制备方法

Country Status (2)

Country Link
CN (1) CN111560044B (zh)
WO (1) WO2021244051A1 (zh)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111560046B (zh) * 2020-06-01 2021-02-05 湖南华诚生物资源股份有限公司 一种作为护肤品抗氧剂的11-o-罗汉果醇肟酯衍生物以及一种护肤品
CN111560044B (zh) * 2020-06-01 2021-02-19 湖南华诚生物资源股份有限公司 一种11-o-罗汉果醇肟醚衍生物及其制备方法

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105832748A (zh) * 2016-05-06 2016-08-10 深圳以诺生物制药有限公司 一种从罗汉果总皂苷中制备罗汉果醇新型衍生物的方法
WO2017075257A2 (en) * 2015-10-29 2017-05-04 Senomyx, Inc. High intensity sweeteners
CN110585053A (zh) * 2019-10-16 2019-12-20 张传旋 用于祛斑、除细纹、黑眼圈和眼袋的复合物及其制备方法
CN111560044A (zh) * 2020-06-01 2020-08-21 湖南华诚生物资源股份有限公司 一种11-o-罗汉果醇肟醚衍生物及其制备方法

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106800580B (zh) * 2017-01-12 2019-05-10 中国药科大学 甾醇类衍生物及其制备方法和应用

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017075257A2 (en) * 2015-10-29 2017-05-04 Senomyx, Inc. High intensity sweeteners
CN105832748A (zh) * 2016-05-06 2016-08-10 深圳以诺生物制药有限公司 一种从罗汉果总皂苷中制备罗汉果醇新型衍生物的方法
CN110585053A (zh) * 2019-10-16 2019-12-20 张传旋 用于祛斑、除细纹、黑眼圈和眼袋的复合物及其制备方法
CN111560044A (zh) * 2020-06-01 2020-08-21 湖南华诚生物资源股份有限公司 一种11-o-罗汉果醇肟醚衍生物及其制备方法

Also Published As

Publication number Publication date
CN111560044B (zh) 2021-02-19
CN111560044A (zh) 2020-08-21

Similar Documents

Publication Publication Date Title
EP3247371B1 (en) Methods of purifying cannabinoids, compositions and kits thereof
WO2021244051A1 (zh) 一种11-o-罗汉果醇肟醚衍生物及其制备方法
US10155708B2 (en) Methods of purifying cannabinoids, compositions and kits thereof
CN101296694A (zh) 药物镓组合物和方法
CN109575099A (zh) 达玛烷皂苷元衍生物及其制备方法和应用
CN111148729A (zh) 苯烯莫德的晶型及其用途与制备方法
CN101434635A (zh) 一类具抗肿瘤活性的水溶性酚性三萜化合物及其制备方法
Luo et al. Water-soluble matrine-type alkaloids with potential anti-neuroinflammatory activities from the seeds of Sophora alopecuroides
EP3252039B1 (en) Compound containing indoleacetic acid core structure and use thereof
WO2014048033A1 (en) Polymorphs of cddo ethyl ester and uses thereof
CN111471080B (zh) ocotillol型人参皂苷元A环并氨基噻唑环衍生物及制备方法
WO2015092720A1 (en) Metabolites of sonidegib (lde225)
KR100406736B1 (ko) 나프토퀴논계 화합물을 포함하는 항암제
CN107056739B (zh) Bola型槲皮素衍生物及其制备方法和应用
AU2021106179A4 (en) New crystalline form of epalrestat as well as preparation method and application thereof
CN109438166A (zh) （1S,2S,4S）-β-榄香烯及其制备方法和应用
CN112745288B (zh) β-烷氧基醇二苯并呫吨类化合物及其应用
Zhou et al. Design, synthesis and anti-tumor activities of carbamate derivatives of cinobufagin
CN107320735A (zh) 一种他莫昔芬组合物及其制剂
CN104402894B (zh) 3-（1-烷氧乙基）二氢卟吩e6类似物的金属络合物及其制备方法和应用
CN102718675A (zh) 阿戈美拉汀甲磺酸复合物及其制备方法
CN114426538B (zh) 一种小檗碱卡格列净衍生物及其制备方法和应用
EP4292593A1 (en) Pharmaceutical composition for preventing cytokine storm
EP4043447A1 (en) Halogenated tetracyclic triterpene derivative, preparation and application thereof
CN103012175A (zh) 抗肿瘤含氮取代基姜黄素类似物、其盐及制备方法

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 21818187

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 21818187

Country of ref document: EP

Kind code of ref document: A1